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中华绒螯蟹 MnGPx-3 和 MnGPx-4 基因的克隆、表达与原位杂交分析及其对低氧和复氧的响应

Molecular cloning, expression, and in situ hybridization analysis of MnGPx-3 and MnGPx-4 from oriental river prawn, Macrobrachium nipponense, in response to hypoxia and reoxygenation.

机构信息

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, Jiangsu, People's Republic of China.

Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, Jiangsu, People's Republic of China.

出版信息

PLoS One. 2020 Feb 21;15(2):e0229171. doi: 10.1371/journal.pone.0229171. eCollection 2020.

Abstract

Glutathione peroxidase (GPx) has been the focus of increased research because of its important role as an antioxidant and in reactive oxygen species (ROS) induced damage repair. Studies on GPxs have relevance with Macrobrachium nipponense because it has poor tolerance to hypoxia in Macrobrachium nipponense. The two subunits named as MnGPx-3 and MnGPx-4 according to the glutathione peroxidase nomenclature system. Both full-length cDNAs were cloned from the hepatopancreas. In this study, we analyzed the expression of two GPxs in Macrobrachium nipponense in response to changes in environmental oxygen. Expression levels of MnGPx-3 and MnGPx-4 indicated that both have strong responses to hypoxia. In situ hybridization showed that MnGPx-3 and MnGPx-4 were located in secretory and storage cells in hepatopancreas. These results suggest that GPx gene is expressed and released by secretory cells and released response to hypoxia. In the gill tissue, however, GPxs are located in blood cells, suggesting that they perform different functions in different tissues or organs. The results of in situ hybridization were consistent with those of quantitative Real-time PCR. This study provides a basis for understanding the oxidative stress response in M. nipponense under hypoxia.

摘要

谷胱甘肽过氧化物酶(GPx)因其作为抗氧化剂和在活性氧(ROS)诱导的损伤修复中的重要作用而成为研究热点。对 GPx 的研究与日本沼虾有关,因为日本沼虾对缺氧的耐受性差。根据谷胱甘肽过氧化物酶命名系统,这两个亚基分别命名为 MnGPx-3 和 MnGPx-4。这两个全长 cDNA 均从肝胰腺中克隆得到。在本研究中,我们分析了两种 GPx 在日本沼虾中对环境氧变化的反应。MnGPx-3 和 MnGPx-4 的表达水平表明它们对缺氧都有强烈的反应。原位杂交显示 MnGPx-3 和 MnGPx-4 位于肝胰腺的分泌和储存细胞中。这些结果表明,GPx 基因由分泌细胞表达和释放,并对缺氧做出释放反应。然而,在鳃组织中,GPx 位于血细胞中,这表明它们在不同的组织或器官中执行不同的功能。原位杂交的结果与定量实时 PCR 的结果一致。本研究为了解日本沼虾在缺氧下的氧化应激反应提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd4f/7034814/05bcaea1863b/pone.0229171.g001.jpg

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